Fast Mass Microscopy: Mass Spectrometry Imaging of a Gigapixel Image in 34 Minutes

Abstract

Mass spectrometry imaging (MSI) maps the spatial distributions of chemicals on surfaces. MSI requires improvements in throughput and spatial resolution, and often one is compromised for the other. In microprobe-mode MSI, improvements in spatial resolution increase the imaging time quadratically, thus limiting the use of high spatial resolution MSI for large areas or sample cohorts and time-sensitive measurements. Here, we bypass this quadratic relationship by combining a Timepix3 detector with a continuously sampling secondary ion mass spectrometry mass microscope. By reconstructing the data into large-field mass images, this new method, fast mass microscopy, enables orders of magnitude higher throughput than conventional MSI albeit yet at lower mass resolution. We acquired submicron, gigapixel images of fingerprints and rat tissue at acquisition speeds of 600,000 and 15,500 pixels s −1 , respectively. For the first image, a comparable microprobe-mode measurement would take more than 2 months, whereas our approach took 33.3 min.

Time-Resolved Imaging of High Mass Proteins and Metastable Fragments Using Matrix-Assisted Laser Desorption/Ionization, Axial Time-of-Flight Mass Spectrometry, and TPX3CAM

“…This reduces the dead time per pixel (475 ns + TOT) and the total readout time of the pixel matrix for occupancies (% of pixel hits) below 50%. ,, This unique data-driven readout system allows the TPX3 to operate at least 10 times faster compared to the previous Ultraflex-TPX experimental setup, where the data is read out by 1 Gbps in both cases . The 10 Gbps ethernet interface at the SPIDR readout system allows the instrument to operate with a higher speed, which would be beneficial in high-throughput mass spectrometry imaging experiments. , …”

“… 11 The 10 Gbps ethernet interface at the SPIDR readout system allows the instrument to operate with a higher speed, which would be beneficial in high-throughput mass spectrometry imaging experiments. 52 , 53 …”

“…11 The 10 Gbps ethernet interface at the SPIDR readout system allows the instrument to operate with a higher speed, which would be beneficial in high-throughput mass spectrometry imaging experiments. 52,53 Further, the single-stop pixels in the TPX limited the detection of higher m/z values, especially at high count rates (e.g., those encountered using MALDI MS) and high MCP voltages that lead to high pixel occupancies, as pixels are deactivated by lower m/z (earlier arriving) ions. In contrast, TPX3 is designed to enable multihit functionality such that each pixel can record multiple events within a single measurement cycle (in this case, for each laser shot).…”

Mass Spectrometry Imaging for Single-Cell or Subcellular Lipidomics: A Review of Recent Advancements and Future Development

“…Microscope mode MSI utilizes a broad laser beam to irradiate the sample in a manner similar to that of a conventional microscope light source. The simultaneous acquisition and preservation of the distribution pattern of specific ions during ionization is a distinguishing aspect of this method, enabling resolution without the limitations imposed by ionization techniques [ 24 , 25 ]. Additionally, the flight path of the ion is monitored to maintain this distribution pattern.…”

Mass spectrometry imaging advances and application in pharmaceutical research

“…Currently, the microprobe-based spot-by-spot or line-by-line scan mode remains the predominant way for MSI data collection, which usually takes hours or even a whole day to complete for a larger size sample or when the ultrahigh-resolution image is required. A rapid, microscopic snapshot-mode MSI data collection will definitely accelerate the MSI working efficiency [116]. This requires a more sophisticated MS instrumentation design in the micro-arrayed ion detection system.…”